LOCUS       sequence-136896-        5123 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NLSPUFa_p65HSF1 in pmax expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5123)
  AUTHORS   Cheng AW, Jillette N, Lee P, Plaskon D, Fujiwara Y, Wang W, 
            Taghbalout A, Wang H
  TITLE     Casilio: a versatile CRISPR-Cas9-Pumilio hybrid for gene regulation 
            and genomic labeling.
  JOURNAL   Cell Res. 2016 Feb;26(2):254-7. doi: 10.1038/cr.2016.3. Epub 2016 
            Jan 15.
  PUBMED    26768771
REFERENCE   2  (bases 1 to 5123)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/cr.2016.3";
            journalName: "Cell Res"; date: "2016-02"; volume: "26"; issue: "2"; 
            pages: "254-7"
COMMENT     Sequence Label: pAC1393-pmax-NLSPUFa_p65HSF1
FEATURES             Location/Qualifiers
     source          1..5123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          813..945
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     protein_bind    960..984
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1002..1011
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1023..1043
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2154..2174
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2190..2732
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse 
                     NF-kappa-B"
                     /label=p65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISS"
     CDS             2757..3128
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     protein_bind    complement(3153..3177)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3230..3271
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3297..3316)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    3326..3407
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3351..3370
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3605..4192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3685..3704)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4273..5067)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4273..5067)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4384..4403)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4994..5013
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagg
      721 tcgtttagtg aaccgtcaga tcactagtag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctc gactgatcac aggtaagtat caaggttaca agacaggttt
      841 aaggaggcca atagaaactg ggcttgtcga gacagagaag attcttgcgt ttctgatagg
      901 cacctattgg tcttactgac atccactttg cctttctctc cacagggaaa aaacaattga
      961 caagtttgta caaaaaagca ggctccgaat tcaccggtgc cgccaccatg gggatcctcc
     1021 cccccaagaa aaagaggaag gtatctagag gccgcagccg ccttttggaa gattttcgaa
     1081 acaaccggta ccccaattta caactgcggg agattgctgg acatataatg gaattttccc
     1141 aagaccagca tgggtccaga ttcattcagc tgaaactgga gcgtgccaca ccagctgagc
     1201 gccagcttgt cttcaatgaa atcctccagg ctgcctacca actcatggtg gatgtgtttg
     1261 gtaattacgt cattcagaag ttctttgaat ttggcagtct tgaacagaag ctggctttgg
     1321 cagaacggat tcgaggccac gtcctgtcat tggcactaca gatgtatggc agccgtgtta
     1381 tcgagaaagc tcttgagttt attccttcag accagcagaa tgagatggtt cgggaactag
     1441 atggccatgt cttgaagtgt gtgaaagatc agaatggcaa tcacgtggtt cagaaatgca
     1501 ttgaatgtgt acagccccag tctttgcaat ttatcatcga tgcgtttaag ggacaggtat
     1561 ttgccttatc cacacatcct tatggctgcc gagtgattca gagaatcctg gagcactgtc
     1621 tccctgacca gacactccct attttagagg agcttcacca gcacacagag cagctggtac
     1681 aggatcaata tggaaattat gtaatccaac atgtactgga gcacggtcgt cctgaggata
     1741 aaagcaaaat tgtagcagaa atccgaggca atgtacttgt attgagtcag cacaaatttg
     1801 caagcaatgt tgtggagaag tgtgttactc acgcctcacg tacggagcgc gctgtgctca
     1861 tcgacgaggt gtgcaccatg aacgacggtc cccacagtgc cttatacacc atgatgaagg
     1921 accagtatgc caactacgtg gtccagaaga tgattgacgt ggcggagcca ggccagcgga
     1981 agatcgtcat gcataagatc cggccccaca tcgcaactct tcgtaagtac acctatggca
     2041 agcacattct ggccaagctg gagaagtact acatgaagaa cggtgttgac ttaggggggc
     2101 cggccggagg aggtggaagc ggaggaggag gaagcggagg aggaggtagc ggacctaaga
     2161 aaaagaggaa ggtggcggcc gctggatccc cttcagggca gatcagcaac caggccctgg
     2221 ctctggcccc tagctccgct ccagtgctgg cccagactat ggtgccctct agtgctatgg
     2281 tgcctctggc ccagccacct gctccagccc ctgtgctgac cccaggacca ccccagtcac
     2341 tgagcgctcc agtgcccaag tctacacagg ccggcgaggg gactctgagt gaagctctgc
     2401 tgcacctgca gttcgacgct gatgaggacc tgggagctct gctggggaac agcaccgatc
     2461 ccggagtgtt cacagatctg gcctccgtgg acaactctga gtttcagcag ctgctgaatc
     2521 agggcgtgtc catgtctcat agtacagccg aaccaatgct gatggagtac cccgaagcca
     2581 ttacccggct ggtgaccggc agccagcggc cccccgaccc cgctccaact cccctgggaa
     2641 ccagcggcct gcctaatggg ctgtccggag atgaagactt ctcaagcatc gctgatatgg
     2701 actttagtgc cctgctgtca cagatttcct ctagtgggca gggaggaggt ggaagcggct
     2761 tcagcgtgga caccagtgcc ctgctggacc tgttcagccc ctcggtgacc gtgcccgaca
     2821 tgagcctgcc tgaccttgac agcagcctgg ccagtatcca agagctcctg tctccccagg
     2881 agccccccag gcctcccgag gcagagaaca gcagcccgga ttcagggaag cagctggtgc
     2941 actacacagc gcagccgctg ttcctgctgg accccggctc cgtggacacc gggagcaacg
     3001 acctgccggt gctgtttgag ctgggagagg gctcctactt ctccgaaggg gacggcttcg
     3061 ccgaggaccc caccatctcc ctgctgacag gctcggagcc tcccaaagcc aaggacccca
     3121 ctgtctccat cgattgatta attaagaatt cgacccagct ttcttgtaca aagtggttga
     3181 tatccagcac agtggcggcc gctcgagtct agagggcccg cggttcgaag gtaagcctat
     3241 ccctaaccct ctcctcggtc tcgattctac gcgtaccggt tagcaattgt tttttcgatg
     3301 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     3361 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     3421 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     3481 acctctacaa atgtggtact taagaggggg agaccaaagg gcgagacgtt aaggcctcac
     3541 gtgacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3601 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3661 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3721 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3781 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3841 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3901 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3961 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4021 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     4081 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacg ctggtagcgg
     4141 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4201 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4261 ggtcatgccg tctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc
     4321 gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc
     4381 agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc
     4441 acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc
     4501 gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc tggcgaacag
     4561 ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc
     4621 ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt
     4681 agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc
     4741 aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc
     4801 ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag
     4861 ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt
     4921 gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc
     4981 gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc
     5041 tgcgtgcaat ccatcttgtt caatcataat attattgaag catttatcag ggttcgtctc
     5101 gtcccggtct cctcccatgc atg
//
